The therapeutic effect of tyrosine kinase inhibitor in 5/6 nephrectomized rats

尼洛替尼是一种第二代酪氨酸酶抑制剂，与伊马替尼相比，它对bcr-Abl的活性提高了30倍，对PDGF受体(PDGFR)和c-Kit的活性与伊马替尼相似，伊马替尼是一种化合物，已被证明在冷球蛋白血症膜增生性肾小球肾炎和肾毒性血清性肾炎等肾脏疾病的动物模型中具有治疗作用。在目前的研究中，我们调查了尼洛替尼在已建立的肾功能衰竭进展中的作用。成年雄性SD大鼠行5/6肾切除或开腹手术(假手术)。5/6肾切除大鼠术后2周开始每日灌胃给予尼洛替尼(45 mg/kg)或赋形剂，共8周。定期测量血压(BP)、尿蛋白(U-P)、血肌酐(Cr)和体重(BW)。处死时进行肾脏形态观察。在体外，我们使用了肾脏…成纤维细胞(NRK49F)和原代系膜细胞较多。尼洛替尼或单独用培养液处理细胞，观察I型胶原合成和PDGFR的变化。用实时定量RT-PCR和免疫印迹法检测血管紧张素II和PDGF-BB诱导的磷酸化。在整个研究过程中，两个治疗组之间的BP和BW具有可比性。治疗6周和8周后，尼洛替尼治疗组大鼠血清肌酐水平显著低于赋形剂组。与赋形剂治疗相比，尼洛替尼治疗的大鼠在治疗后1周表现出U-P降低。在整个研究过程中，这种减少一直保持不变。尼洛替尼治疗后，除了肾小球硬化和肾小管间质损害评分降低外，残余肾肥大程度也有所减轻。尼洛替尼治疗组肾皮质I型胶原、转化生长因子-β、纤维连接蛋白和纤溶酶原激活物-1的表达也显著降低。在体外，尼洛替尼阻断血管紧张素-II诱导的肾成纤维细胞和肾小球系膜细胞I型胶原/GAPDHmRNA的产生。尼洛替尼还可降低I型胶原/GAPDHmRNA水平，阻止PDGFR。PDGF-BB诱导系膜细胞磷酸化。尼洛替尼治疗可显著减轻体内和体外的肾脏纤维化。我们的结果表明，尼洛替尼在限制慢性肾脏疾病进展到终末期肾功能衰竭方面可能是有用的。较少

Nilotinib is a second-generation tyrosine kinase inhibitor that demonstrates a 30-fold increase in activity against Bcr-Abl, and a similar level of activity against the PDGF receptor(PDGFR) and c-Kit when compared to imatinib, a compound that has been previously shown to exhibit therapeutic benefits in animal models of renal disease, including cryoglobulinemic membranoproliferative glomerulonephritis and nephrotoxic serum nephritis. In the current study, we investigated the role of nilotinib in the progression of established renal failure. Adult male Sprague Dawley rats were subjected to 5/6 nephrectomy or laparotomy(sham-operated). Rats with 5/6 nephrectomy were then administered either nilotinib(45mg/kg) or vehicle via daily oral gavage from 2 weeks after surgery, and for a period of 8 weeks. Blood pressure(BP), proteinuria(U-P), serum creatinine(Cr) and body weight(BW) were measured periodically. Renal morphological investigations were performed at sacrifice. In vitro, we used renal … More fibroblasts(NRK49F) and primary mesangial cells. Cells were pretreated with nilotinib or medium alone, and collagen type I synthesis and PDGFR. phosphorylation induced by angiotensin II or PDGF-BB were analyzed by real-time RT-PCR and immunoblotting. BP and BW were comparable between the two treatment groups throughout the study. Following 6 and 8 weeks of treatment, serum Cr levels in the nilotinib-treated rats were significantly lower than that of the vehicle-treated rats. When compared to vehicle treatment, nilotinib-treated rats demonstrated reduced U-P at 1 week after treatment. This reduction was maintained over the course of the study. Nilotinib treatment also resulted in a decrease in remnant kidney hypertrophy, in addition to reduced scores of glomerulosclerosis and tubulointerstitial damage. Renal cortical mRNA for collagen type I, TGF-β, fibronectin, and PAI-1 were also significantly decreased in the nilotinib treated group. In vitro, nilotinib blocked collagen type I/GAPDH mRNA production induced by angiotensin-II in renal fibroblasts and mesangial cells. Nilotinib also decreased collagen type I/GAPDH mRNA levels and prevented PDGFR. phosphorylation induced by PDGF-BB in mesangial cells. Nilotinib treatment significantly attenuates renal fibrosis in vivo and in vitro. Our results suggest that nilotinib may prove useful in limiting the progression of chronic renal disease to end-stage renal failure. Less